에틸렌옥사이드에 폭로된 흰쥐의 혈액에 형성된 헤모글로빈 부가체에 대한 연구

이진헌,신호상,안혜실,Lee Jin-Heon,Shin Ho-Sang,Ahn Hye-Sil 한국환경보건학회 2006 한국환경보건학회지 Vol.32 No.2

Ethylene oxide is a genotoxic carcinogen with widespread uses as industrial chemical intermediate and gaseous sterilant. 2-hydroxyethylated N-terminal valine in Hb is a good biomarker for biological monitoring of ethylene oxide exposure, because of its stability. For measuring the hemoglobin adduct formed by exposure of ethylene oxide, we studied the determination of (N-2-hydroxy-ethyl)valine(HEV) in hemoglobin adduct by using GC/MS. Firstly we synthesized HEV with 2-amino-ethanol and bromoisovaleric acid(BIVA) and confirmed it with GC/MS-FID. Its fragmentations were m/z 116(base ion, M+-45) and m/z 130(M+-31). For measuring HEV with higher sensitivity, we use derivatives which were PFPITH(pentafluorophenylisothiocianate) and TBDMS (tributyldimethylsilylation) by using Edman procedure. Its fragmentation were m/z 425(M+-57), m/z 383(M+-99) and m/z 172(M+-310) by using GC/MS. We did biological monitoring for mice inhalation exposure with 400 ppm ethylene oxide. The concentrations of hemoglobin adduct were $168{\pm}3.8\;and\;512{\pm}04$(nmol g-1 globin) at 0.5 hr/day 400 ppm ethylene oxide inhalation exposure group, and $631{\pm}17\;and\;2265{\pm}9.4$(nmol g-1 globin) at 1.0 hr/day 400 ppm ethylene oxide inhalation exposure for 1 and 4 weeks, respectively. We confirmed that (N-2-hydroxy-ethyl)valine(HEV) of hemoglobin was a good biomarker for biomonitoring of ethylene oxide exposure, and can measured with derivatives such as PFPITH(pentafluorophenylisothiocianate) and TBDMS(tributyldimethylsilylation) by using GC/MS.

근로자의 뇨중 상피세포에서 ³²P-postlabeling에 의한 발암물질의 DNA adducts측정방법에 대한 연구

이진헌,노재훈,그린 탈라스카,Lee, Jin Heon,Roh, Jaehoon,Talaska, Glenn 한국산업보건학회 2000 한국산업보건학회지 Vol.10 No.1

Carcinogen-DNA adduct analysis has potential for biomonitoring the earliest effects of exposure to many chemical carcinogens. They are the covalent reaction products of electrophiles and nucleophilic sites on DNA and the initial damage to DNA induced by many carcinogens. So many researchers begin to use them as biomarker for monitoring the earliest exposure of carcinogens and develop the effective analytical techniques about them. Randerath, Gupta and coworkers(1981, 1982) has also developed a $^{32}P$-postlabelling method as one among them. A major project for biomonitoring workers with carcinogen-DNA adducts is to develop non-invasive samples instead of tissues of target organs such as baldder and lung. This study use the exfoliated urothelial cells in urine for examine benzidine-DNA adducts. The content of exfoliated urothelial cells is not enough to significantly measure DNA content with spectrophotometer, and require the another way. So firstly washing the collected cells with PBS and 70% ethanol and centrifuge them for removing the crystals in urine, which block the isolation of DNA adducts. And then, measure the total nucleotide after $^{32}P$-postlabelling for calculating RAL. $[{\gamma}-^{32}P]ATP$ using for $^{32}P$-postlabelling, can synthesize with $[^{32}P]H_3PO_4$, and reagent and enzyme mixture (RM, EM), which is very economic in case of requiring a lot of them. Chromatography was composed of two steps. First step was to separate adduct ones from unadducted nucleotide, and secondary step was separate each adduct, which were performed with 4 kinds of solvents and different directions on TLC. With this procedure, we measure the DNA adducts in exfoliated urothelial cells of workers who were employed in benzidine and benzidine-dye company. RAL of adducts were $89.0{\times}10^7$ and $57.0{\times}10^7$ in them. In conclusion, we can significantly measure the DNA adduct in exfoliated urothelial cells by using the above $^{32}P$-postlabelling procedures, and use them to be biomonitoring workers who exposed carcinogens.

디클로로벤지딘에 폭로된 흰쥐의 간장세포와 방광 상피세포에 형성된 DNA adducts의 $^{32}$ P-postlabeling과 GC/MS-SIM에 의한 분석

이진헌,신호상,장미선,Lee Jin Heon,Shin Ho-Sang,Jang Mi Seon 한국환경보건학회 2002 한국환경보건학회지 Vol.28 No.1

To identify and evaluate the dichlorobenzidine(DCB)-DNA adducts in liver cell and bladder epithelial cells by $^{32}$ P-postlabeling and GC/MS-SIM, we orally exposed the dichlorobenzidine(20mg/kh body wt./day) to male Sprague-Dawley rats(l85$\pm$10g) for 14 days. Two kinds of DCB-DNA adduct(A1 and A2) were found at the same site of thin layer chromatogram of $^{32}$ P-postlabeling method in liver cells and bladder epithelial cells. In liver cells, relative adduct labeling(RAL) $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 34.1$\pm$3.71 and 69.9$\pm$5.02, that of adduct A2 were 74.1$\pm$10.1 and 105.1$\pm$10.1 on 10 and 14 days after treatment, respectively. And in bladder epithelia cells, RAL $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 5.92$\pm$1.60 and 15.9$\pm$1.31, that of adduct A2 were 9.81$\pm$2.81 and 22.8$\pm$1.79 on 10 and 14 days after treatment, respectively. DCB metabolites formed DNA adducts were monoacetyl-dichlorobenzidine(acDCB) and diacetyl-dichlorobenzidine(di-acDCB), which was identify by gas chromatography/mass spectrometry-scan ionization mode(GC/MS-SIM), after hydrolysis of DCB-DNA adducts isolated from live cells and bladder epithelial cells. The base peak of acDCB were 252 and 294 m/z, and that of di-acDCB were 252, 294 and 336 m/z. In conclusion, the exposed DCB formed two kinds of DCB-DNA adduct, the proximate materials of that were acDCB and di-acDCB in liver and bladder epithelial cells. And the above GC/MS-SIM method was found the DCB-DNA adducts could be monitoring by gas chromatography.

한ㆍ미ㆍ일 시나리오 구성 비교

이진헌,Lee, Jin-Heon 한국데이터베이스진흥원 2006 디지털콘텐츠 Vol.1 No.-

오늘날 독자들을 매료시키고 있는 디지털 문학이란 과연 19세기 소설처럼 온화하고 책임감 있는 것일까, 아니면 환각 작용을 일으키는 마약처럼 위험스럽고 치명적인 것일까, 현재 우리는 뉴미디어 혁명의 한가운데에서 있다. 14세기의 인쇄 활자나 19세기의 사진 기술이 당대의 사회와 문화에 혁명적인 충격을 주었던것처럼, 이제는 모든 문화가 컴퓨터를 매체로 생산 ㆍ배포ㆍ의사소통하고 있다. 본고에서는 헐리우드 공식을 기반으로 한ㆍ미ㆍ일 애니메이션 시나리오 구성 방식으 비교했다.

유기수은 노출로 인한 흰쥐 태자골격성장 지연에 대한 마늘의 보호효과

이진헌,박경렬,김대선,Lee, Jin-Heon,Park, Kyung-Lyoul,Kim, Dae-Seon 한국환경보건학회 2008 한국환경보건학회지 Vol.34 No.2

This study evaluates the protective effects of Korean garlic juice against the toxicity of methyl mercuric chloride (MMC) in relation to fetal ossification in pregnant Fischer 344 rats. This study has as its basis, both theory and data that neutral amino acids in garlic juice have protective effects against mercury poisoning. Pregnant rats were dosed in various combination with 20 mg MMC/kg body wt. and 0.5 or 1.0 garlic juice/kg body wt. on the 7th day of gestation. Fetuses were extracted from the dams on 20th day of gestation, and the fetal bones were stained and measured. The results obtained are as follows: 1. Fetal body weights, body lengths, and head length were significantly decreased by as much as 23.5%, 21.3%, 15.5% respectively in 20 mg/wt kg methyl mercuric chloride groups (p<0.05). But in the garlic treated groups, they were almost similar to the controls. 2. The ossification centers were significantly decreased by as much as 35.1% in pelvic phalanges, 53.5% in pectoral phalanges, 74.1% in ternebrae, 76.2% in tail in 20 mg/wt kg methyl mercuric chloride groups (p<0.05). But in the garlic treated groups, they were significantly increased by as much as $81.2{\sim}88.7%$ and $94.8{\sim}98.9%$ of controls. 3. The ossified pectoral girdles were significantly decreased by as much as 66.6% in clavicle, and in other areas $74.2{\sim}87.4%$ in the 20 mg/wt kg methyl mercuric chloride groups (p<0.05). But in the garlic treated groups, they were significantly increased by as much as $81.2{\sim}88.7%$ and $94.8{\sim}98.9%$ of the controls. 4. The ossified pelvic girdles were significantly decreased by as much as 57.1% and 56.1% in two ischium, $67.2{\sim}81.7%$ in metacarpals in the 20 mg/wt kg methyl mercuric chloride groups (p<0.05). But in the garlic treated groups, they were significantly increased by as much as $67.0{\sim}85.6%$ and $90.1{\sim}98.7%$ of the controls. 5. The ossified terenebrae were unchanged or significantly decreased; 0.0% in 5th, 54.1% in 1st, 83.9% in 2nd, 75.0% in 3rd, 72.7% in 4th, 79.8% in 6th of 20 mg/wt kg methyl mercuric chloride groups (p<0.05). But in the garlic treated groups, they were significantly increased by as much as $29.5{\sim}55.1%$ and $54.5{\sim}84.0%$ of the controls. 6. The ossified ribs were significantly decreased by as much as $8.3{\sim}18.0%$ in 20 mg/wt kg methyl mercuric chloride groups (p<0.05). But in the garlic treated groups, they were significantly increased by as much as $87.1{\sim}93.5%$ and $96.3{\sim}99.7%$ of the controls. In conclusion, Korean garlic juice significantly protected against the toxicity of MMC in relation to the fetal ossification in pregnant rats.

병아리 태자에 대한 엔로플록사신의 최기형 독성 연구

이진헌,이지희,조이연,Lee, Jin-Heon,Lee, Ji-Hee,Cho, Y-Eaun 한국환경보건학회 2008 한국환경보건학회지 Vol.34 No.4

The purpose of this study was to investigate the teratological effects of enrofloxacin, a veterinary antibiotic, on the embryos and fetus of hatching chicken eggs. A control group and four experimental fertilized egg groups were set up. The four experimental groups were injected with 0.05 ml, 0.1 ml, 0.2 ml, and 0.3 ml enrofloxacin ($50{\mu}g/ml$) respectively. During the hatching period, the weights of total eggs, inside material(yolk and white) and embryos or fetuses, embryo growth, and teratological effects were investigated. In the experimental groups (fertilized eggs injected with 0.05 ml, 0.1 ml, 0.2 ml and 0.3 ml enrofloxacin), the weights of total eggs were decreased, but the yolk and white weights were increased by the higher amount of antibiotics. The weights of embryos (fetuses) in experimental groups were 86.1%, 78.6%, 65.6% and 61.4% of the control, respectively. Retarded growth, deformity and embryo loss were observed in experimental groups. Teratological effects such as undeveloped eyes, wings and legs, and deformed head and bill were also detected. In conclusion, we found that veterinary antibiotics, enrofloxacin have made teratological effects on the embryos and fetus of hatching eggs.

한국 어린이와 청소년의 요중 크레아티닌 농도와 영향요인에 대한 연구

이진헌,안령미,강희숙,최석남,홍춘표,김진경,Lee, Jin-Heon,Ahn, Ryeong-Mi,Kang, Hee-Sook,Choi, Suk-Nam,Hong, Chun-Pyo,Kim, Jin-Gyong 한국환경보건학회 2012 한국환경보건학회지 Vol.38 No.4

Creatinine-adjustment is an important process in the urinary monitoring of the environmental exposure of children and adolescents. The purpose of this study was to investigate the concentrations of urinary creatinine and factors associated with them among Korean children and adolescents. We recruited 1,025 persons from 128 extracted schools. They were from three to 18 years old and supplied urine samples for measuring creatinine. The concentrations of urinary creatinine were 98.18 mg/dl (SD, 67.67) in arithmetic mean and 72.05 mg/dl (GSD 2.49) in geometric mean, were significantly higher among male children/adolescents than females in all age groups, and higher values appeared following increasing ages, heights and BMIs. The rates of the number who were below the lowest limit recommended by WHO (<30 mg/dl) were 25.57% among three to four year olds, 21.77% among five to six year olds, 20.0% among seven to eight year olds and 14.69% among nine to ten year olds, respectively. The rates of those above the highest limit (>300 mg/dl) were 0.0% among three to twelve year olds. The coefficient of determination R-square of the fitted regression model for urinary creatinine was 27.4% with general characteristic variables of sex, age, BMI and height. The significant variables among these were height (standardized beta = 0.372) and age (standardized beta = 0.129). Another coefficient of determination R-square was 15.3% with dietary habit variables of smoking, drinking, dining area, number of meals and snacks, and intake of milk food, cup-noodles, canned foods, popcorn, nachos, and hamburgers. In conclusion, the concentration of urinary creatinine was significantly lower in children than in adults, and was very significantly associated with the height of children. Therefore, children need the recommended concentrations for urinary creatinine, as distinguished from adults.

한국 성인의 요중 크레아티닌 농도 변화에 대한 성, 연령 그리고 체질량지수(BMI)의 관련성 연구

이진헌,안령미,Lee, Jin-Heon,Ahn, Ryoung-Me 한국환경보건학회 2010 한국환경보건학회지 Vol.36 No.3

The purpose of this study was to investigate the relevance of gender, age, and BMI (Body Mass Index) to changes in the urinary creatinine concentration in Korean adults. We recruited and surveyed 2,156 persons $\geq$20 years of age from 98 districts across the country, and collected urine samples for analyzing the creatinine concentration. Participants were 41.6% men and 58.4% women. In terms of age, the percentage of the population in their 20's and $\geq$60 years of age was relatively similar, with 13.6% and 17.8%, respectively, while the percentages in their 30's, 40's and 50's were also relatively similar, with 22.0%, 24.0%, and 22.6%, respectively. The proportion of participants in their 20's and 30's who were underweight (according to the BMI) was 25% and 19.2%, respectively, but only 6.2~6.4% of those $\geq$40 years of age were underweight. This showed that the underweight proportion in elder generations was less than that in younger generations. Meanwhile, the proportion who were overweight was 13.7%, 18.8%, 28.6%, 27.9%, 32.0% when the participants were in their 20's, 30's, 40's, 50's and $\geq$60 years of age, respectively (p<0.000). This showed that the overweight proportion increased along with increasing age. The overall urinary creatinine concentration was 10 mg/dl (arithmetic mean: AM), and 92.2 mg/dl (geometric mean: GM). The urinary creatinine concentration in men (132.6 mg/dl, AM) was significantly higher than that in women (93.3 mg/dl, AM (p<0.000)). Showing a similar trend in men and women, urinary creatinine concentrations were highest when the participants were in their 20's (135.6 mg/dl, AM), and tended to decrease with increasing age. Urinary creatinine concentrations in overweight and obese subjects (AM of 117.9 mg/dl and 118.0 mg/dl, respectively) were significantly higher than in other groups, and this trend was similar in men and women. In conclusion, we found that urinary concentrations were significantly affected by gender, age, and BMI, and that care should therefore be exercised when correcting urinary metabolites according to the urinary creatinine concentration.

GC-MS-SIM과 ³²P-postlabeling방법에 의한 염료와 안료를 제조하는 사업장 근로자들의 방광암 조기발견을 위한 Biomarker개발에 관한 연구

이진헌(Jin Heon Lee),신호상(Ho Sang Shin),김양호(Yang Ho Kim),노재훈(Jae Hoon Roh),이범규(Beom Gyu Lee) 한국환경보건학회 2000 한국환경보건학회지 Vol.26 No.1

To develop the biomarker of non-invasive biomonitoring among workers who exposed benzidine-based dyes in its manufacturing and treating industries, we examined and evaluated the specific carcinogen-DNA adducts in exfoliated urothelial cells of 33 exposed workers. Samples were collected from urine of workers, and then isolated DNA from them. DNA adducts were analyzed using ³²P-postlabeling, along with Nuclease P₁. One adducts was predominated in those samples with 6 adducts present. It was formed in all exposed workers, but the others not. Its relative adducts level(RAL) was 73.2 × 10⁷, which was statistically significantly higher than any other adducts levels(p<0.05) . Analyzing regression between its RAL and employment period of workers, simple linear regression was positively increased with slop of 6.922(p<0.05, r²=0.548). And also, DNA adducts were analyzed using GC/MS-SIM, along with n-butanol extraction and trifluoroacetyl anhydride(TFA) derivatization. Standard material was synthetic N( deoxyguanosin-8-yl)-N -acetylbenzidine. The characteristics of sample and standard were observed on the same sites, such as M⁺(m/z 376) and M⁺(m/z 279) for benzidine-di-TFA and mono-TFA. And then, they were confirmed by analyzing with secondary ion mass(SIM). This study showed that benzidine-based dyes was formed one DNA adduct into significantly high levels in bladder of all exposed workers, its chemical formula was N-(deoxyguanosin-8-yl)-N acetylbenzidine. So we can do monitoring the workers with exfoliated urothelial cells of them by analyzing above DNA adduct as biomarker for preventing bladder cancer causing the exposure of the benzidine-based dyes.

1,3-부타디엔 호흡기 노출의 생체지표 대사물질에 대한 연구

이진헌 ( Jin Heon Lee ) 한국산업위생학회 2010 한국산업보건학회지 Vol.20 No.1